Sperm Release Research Articles

Myosin VI Is Associated With the Endoplasmic Reticulum in Regions of Sertoli Cells Containing Tubulobulbar Complexes.

Myosin VI has been reported by others to localize in association with various regions of apical tubulobulbar complexes (TBCs) at sites of attachment between Sertoli cells and late spermatids in the mouse. Tubulobulbar complexes internalize "intact" intercellular junctions during sperm release and during spermatocyte translocation through the blood-testis barrier. Here, we use super-resolution (STED-stimulated emission depletion) and electron microscopy of immunolabeled sections of rat testis to clearly define the localization of anti-myosin VI reactivity both at apical and basal sites in the epithelium. In data stacks collected by STED imaging, staining at TBCs was predominantly associated with bulb regions of the complexes. At apical sites, when data stacks were analyzed with an Imaris software, staining appeared around and extended between adjacent bulbs. At basal sites, in addition to labeling at TBC bulbs, reactive sites appeared concentrated in regions close to but not directly associated with intercellular junctions. At the ultrastructural level, labeling was predominantly associated with cisternae of the endoplasmic reticulum associated with the bulbs of TBCs and near to basal junction complexes. We conclude that myosin VI may be associated with specific subdomains of the endoplasmic reticulum related to TBC bulbs and associated basal junction complexes between Sertoli cells.

The stony coral Fimbriaphyllia (Euphyllia) ancora’s reproductive strategy involves a sex change every year

A sex change phenomenon was reported in some free-living, non-sessile coral species of the Family Fungiidae. However, there are no reports describing sex change in sessile colonial species. Timing and cellular processes of sex change are also unclear in corals. Here, we report sex change of the colonial coral, Fimbriaphyllia ancora, and its cellular process. Of 26 colonies monitored at Nanwan Bay, southern Taiwan, about 70% changed their sex every year after annual spawning for least 3-4 consecutive years, i.e., colonies that were male two years ago became female last year, and male again this year. The remaining 30% were permanently male or female. Sex-change and non-sex-change colonies grew in close proximity or even side-by-side. No significant differences were found in colony size between sex-change and non-sex-change colonies. Histological analysis showed that, in female-to-male sex change, small oocytes were present up to 3 months in some gonads after spawning and disappeared by 5 months. This suggests that sex change occurred 4-5 months after spawning. In contrast, in male-to-female sex change, oocytes appeared weeks after sperm release and in most gonads by 3 months, suggesting that male-to-female sex change occurred 0–3 months after sperm release.

Gametogenesis and seminatural reproduction of the Amazon twospot astyanax Astyanax bimaculatus (Linnaeus, 1758) cultivated in an enriched environment

Environmental enrichment is used to provide well-being to the animals, such as fish, in captive conditions, mimicking their natural habitat. It may influence fish behavior, physiology, and survival. In terms of reproduction, however, the relationship between environment enrichment and successful reproduction in captivity is still poorly explored in fish species. Aiming to understand any possible benefits of structural enrichment on fish reproduction, 10-days-hatched larvae of the twospot astyanax Astyanax bimaculatus were raised for 18 weeks in tanks with different elements of structural environmental enrichment (PVC pipes, stones, and artificial plants). In the 5th month of life, those animals were hormonally induced to reproduce to assess gamete formation and offspring quality. Animals raised in a sterile-reared environment (non-enriched) showed earlier spawning than the enriched one, presenting significant quantities of Postovulatory follicle complexes (POCs) and cells in atresia in female ovaries, indicating possible reproductive dysfunction or stress, as well as a greater quantity of empty testicular lumen in males, indicating great release of sperm. On the contrary, animals cultivated in enriched environments showed gonads filled with semen in males and vitellogenic oocytes in females. Furthermore, offspring from the sterile-reared group presented significant rates of larval abnormality compared to the enriched group. In conclusion, the results of this study show that environmental enrichment can interfere with the reproduction of fish in captivity, mainly by preventing early maturation of gametes, which can result in low-quality offspring and, consequently, low production of fish species.

MpMLO1 controls sperm discharge in liverwort.

In bryophytes, sexual reproduction necessitates the release of motile sperm cells from a gametophyte into the environment. Since 1856, this process, particularly in liverworts, has been known to depend on water. However, the molecular mechanism underlying this phenomenon has remained elusive. Here we identify the plasma membrane protein MpMLO1 in Marchantia polymorpha, a model liverwort, as critical for sperm discharge from antheridia. The MpMLO1-expressing tip cells among the sperm-wrapping jacket cells undergo programmed cell death upon antheridium maturation to facilitate sperm discharge after the application of water and even hypertonic solutions. The absence of MpMLO1 leads to reduced cytoplasmic Ca2+ levels in tip cells, preventing cell death and consequently sperm discharge. Our findings reveal that MpMLO1-mediated programmed cell death in antheridial tip cells, regulated by cytosolic Ca2+ dynamics, is essential for sperm release, elucidating a key mechanism in bryophyte sexual reproduction and providing insights into terrestrial plant evolution.

The surprising complexity and diversity of sperm storage structures across Galliformes.

In internal fertilisers, the precise timing of ovulation with the arrival of sperm at the site of fertilisation is essential for fertilisation success. In birds, mating is often not synchronised with ovulation, but instead females utilise specialised sperm storage tubules (SSTs) in the reproductive tract, which can ensure sperm are always available for fertilisation at the time of ovulation, whilst simultaneously providing a mechanism of post-copulatory sexual selection. Despite the clear importance of SSTs for fertilisation success, we know little about the mechanisms involved in sperm acceptance, storage, and release. Furthermore, most research has been conducted on only a small number of species, based on which SSTs are usually assumed to look and function in the same way across all species. Here, we conduct a comparative exploration of SST morphology across 26 species of Galliformes. We show that SSTs, and the surrounding tissue, can vary significantly in morphology across species. We provide observational evidence that Galliformes exhibit at least 5 distinct categories of tubule types, including distinctive coiled and multi-branched tubules, and describe 2 additional features of the surrounding tissue. We suggest functional explanations for variation in tubule morphology and propose next steps for future research. Our findings indicate that SSTs are likely to be far more variable than has previously been assumed, with potentially important consequences for our understanding of sperm storage in birds and post-copulatory sexual selection in general.

Male allocation to ejaculation and mating effort imposes different life history trade-offs.

When males compete, sexual selection favors reproductive traits that increase their mating or fertilization success (pre- and postcopulatory sexual selection). It is assumed that males face a trade-off between these 2 types of sexual traits because they both draw from the same pool of resources. Consequently, allocation into mate acquisition or ejaculation should create similar trade-offs with other key life history traits. Tests of these assumptions are exceedingly rare. Males only ejaculate after they mate, and the costs of ejaculation are therefore highly confounded with those of mating effort. Consequently, little is known about how each component of reproductive allocation affects a male's future performance. Here, we ran an experiment using a novel technique to distinguish the life history costs of mating effort and ejaculation for mosquitofish (Gambusia holbrooki). We compared manipulated males (mate without ejaculation), control males (mate and ejaculate), and naïve males (neither mate nor ejaculate) continuously housed with a female and 2 rival males. We assessed their growth, somatic maintenance, mating and fighting behavior, and sperm traits after 8 and 16 weeks. Past mating effort significantly lowered a male's future mating effort and growth, but not his sperm production, while past sperm release significantly lowered a male's future ejaculate quantity, but not his mating effort. Immune response was the only trait impacted by both past mating effort and past ejaculation. These findings challenge the assumption that male reproductive allocation draws from a common pool of resources to generate similar life history costs later in life. Instead, we provide clear evidence that allocation into traits under pre- and postcopulatory sexual selection have different trait-specific effects on subsequent male reproductive performance.

Allochronic reproductive cycles among colonies of the Caribbean octocoral Antillogorgia americana

AbstractThe reproductive biology of the branching octocoral Antillogorgia americana was studied at a site on the Caribbean coast of Panama in 1990–1991 by examining the reproductive status of 11 colonies across 14 months. Colonies were gonochoric. The presence of large and mature eggs or spermaries was allochronic across colonies and months, with peak gonad volumes occurring in months ranging from October through May. Reproductive effort varied between branches on a colony, with variation between branches and branchlets accounting for 25% of the random variation between polyps. Branchlets at the tip of the colony had fewer mature eggs than those lower on the branch, and polyps at the tips of the branchlets had fewer still. Although the simultaneous release of eggs and sperm is critical to reproductive success, the lack of synchrony among colonies on the scale of months may reflect less need for all colonies to spawn in a single event among abundant species that release large numbers of gametes. Such a strategy also spreads the risk of reproductive failure due to environmental conditions during any single month. The presence of multiple spawning episodes can also drive the reproductive isolation of populations and may reflect the presence of cryptic species within the taxon. Studies of reproductive timing can be an important adjunct in identifying variation in life history strategies as well as assessing the validity of species boundaries.

Retinoic acid-releasing scaffold based on chitosan hydrogel and testis decellular plates

Introduction: The use of releasing scaffolds is promising for testes tissue engineering. Chitosan (CS) is a natural biopolymer extensively used as a delivery system. The decellularized testis provides a structure resembling natural extracellular matrix (ECM). All-trans retinoic acid (atRA) is an important factor for spermatogonia differentiation, meiosis completion, and mature sperm release. In this study, thermosensitive CS/βGP hydrogel was served as a novel atRA-releasing support for testis decellular plates (TDPs). Methods: The CS/βGP hydrogel was evaluated for gelation time, morphology, wettability, cytocompatibility, and atRA-releasing behavior. Mouse testes were treated with 1% SDS and evaluated for decellularization efficacy through morphological assessments, DNA content assays, and DAPI staining. TDPs were obtained from the decellularized testes and placed on an atRA-releasing CS/βGP hydrogel support. Results: The CS/βGP hydrogels were prepared with different formulations. It was found that increasing the βGP concentration significantly decreased the gelation time. The addition of atRA did not considerably affect the hydrophilicity of hydrogel. The in vitro release studies showed a sustained atRA release behavior, although an initial low burst release was recorded. Also, increasing the amount of atRA led to a decrease in the rate of drug release. The decellularization procedure successfully removed cells while preserving the ECM. The atRA-releasing CS-TDP scaffold was found to be non-toxic with good biocompatibility. Conclusion: Results showed that the novel atRA-releasing CS-TDP scaffold can sustainably deliver atRA to the culture system and create a cytocompatible environment for testicular cells. Therefore, this scaffold may be useful in developing new tissue engineering approaches for various types of male infertility diseases.

Let's talk about sex: Mechanisms of neural sexual differentiation in Bilateria.

In multicellular organisms, sexed gonads have evolved that facilitate release of sperm versus eggs, and bilaterian animals purposefully combine their gametes via mating behaviors. Distinct neural circuits have evolved that control these physically different mating events for animals producing eggs from ovaries versus sperm from testis. In this review, we will describe the developmental mechanisms that sexually differentiate neural circuits across three major clades of bilaterian animals-Ecdysozoa, Deuterosomia, and Lophotrochozoa. While many of the mechanisms inducing somatic and neuronal sex differentiation across these diverse organisms are clade-specific rather than evolutionarily conserved, we develop a common framework for considering the developmental logic of these events and the types of neuronal differences that produce sex-differentiated behaviors. This article is categorized under: Congenital Diseases > Stem Cells and Development Neurological Diseases > Stem Cells and Development.

Multiple ageing effects on testicular/epididymal germ cells lead to decreased male fertility in mice

In mammals, females undergo reproductive cessation with age, whereas male fertility gradually declines but persists almost throughout life. However, the detailed effects of ageing on germ cells during and after spermatogenesis, in the testis and epididymis, respectively, remain unclear. Here we comprehensively examined the in vivo male fertility and the overall organization of the testis and epididymis with age, focusing on spermatogenesis, and sperm function and fertility, in mice. We first found that in vivo male fertility decreased with age, which is independent of mating behaviors and testosterone levels. Second, overall sperm production in aged testes was decreased; about 20% of seminiferous tubules showed abnormalities such as germ cell depletion, sperm release failure, and perturbed germ cell associations, and the remaining 80% of tubules contained lower number of germ cells because of decreased proliferation of spermatogonia. Further, the spermatozoa in aged epididymides exhibited decreased total cell numbers, abnormal morphology/structure, decreased motility, and DNA damage, resulting in low fertilizing and developmental rates. We conclude that these multiple ageing effects on germ cells lead to decreased in vivo male fertility. Our present findings are useful to better understand the basic mechanism behind the ageing effect on male fertility in mammals including humans.

Genes associated with cell modelling provides new insights into spermiation mechanism in Cyprinus carpio

Spermiation, an act of sperm release, depends on several molecular factors. Despite hormonal administration, spermiation failure is a primary concern in certain fishes. In this study, the molecular mechanisms of spermiation have been analyzed in Cyprinus carpio by comparative transcriptomics. Unigenes for C. carpio control (CCC), which were injected with PBS (Phosphate-buffered saline), and C. carpio treated (CCT), which were injected with ovatide, were 107,616 and 133,435, respectively. A total of 93 genes were identified as involved in the spermiation process, including those related to gonadal steroidogenesis, cell growth, cell adhesion, and cytoplasmic matrix formation. The cd63, CENPS, rasa1a, and genes for gonad steroidogenesis, cell growth, cell adhesion, and cytoplasmic matrix formation were analyzed. Gene expression analysis revealed tubulobulbar complexes mediated disengagement of spermatozoa and JAK2 signaling regulated cyst breakage in teleost for the first time. Analysis was done from the changes at the molecular level to the final act of spermiation. Tissue histology analysis was conducted in accordance with the molecular study, which showed structural changes. Induced breeding in fish plays a key role in seed production in aquaculture sector. However, there are several constraints the sector is still facing due to lack of extensive knowledge regarding the mechanisms of spermiation and species-specific response to hormonal dosage. This study is relevant to understand the molecular mechanisms involved in spermiation and the stages which mark as critical point of sperm release after administrating the inducing agent. This study also lays the groundwork for further exploration of species-specific responses to hormonal treatments, aiding sustainable seed production in the fisheries sector.

Semen collection and characterization of normative reproductive traits in free-ranging ocelots (Leopardus pardalis) and bobcats (Lynx rufus) in southern Texas

Decreased genetic diversity and possible inbreeding depression have recently been documented in the last wild ocelot (Leopardus pardalis) population in the United States. One consequence of inbreeding depression in felids may be reduced semen quality which can adversely affect reproductive potential. Detailed assessments of reproductive parameters in wild individuals and populations can be conducted using assisted reproductive technologies, such as semen collection and analysis. For most felid species, semen has traditionally been collected via electroejaculation (EEJ22Electroejaculation); however, an alternative method has been developed using alpha-2 agonist drugs to induce direct sperm release into the urethra, allowing collection by catheterization without requiring specialized equipment. The goal of this study was to characterize normative reproductive traits in free-ranging ocelots and co-occurring bobcats (Lynx rufus) in southern Texas and assess the effectiveness of urethral catheterization (UC33Urethral Catheterization) for semen recovery in both species. For semen collection, free-ranging cats were live-captured and anesthetized using intramuscular ketamine and medetomidine/dexmedetomidine (alpha-2 agonist) with UC conducted 20–40 minutes post-induction. In ocelots only, EEJ was subsequently performed if UC failed to recover a viable sample. Semen collection was attempted in 31 felids (n=9 ocelots; n=22 bobcats), with sperm recovery by UC in seven of nine ocelots (78 %) and 14 of 22 bobcats (66 %), and by EEJ in four of five ocelots (80 %). For ocelots, the percentage of primary morphologic abnormalities was higher (p<0.001) for UC (47.75 ± 6.7; mean ± SEM) compared to EEJ (9 ± 2.7) but percent normal morphology (MORPH) did not differ between UC and EEJ (p=0.218). In wild ocelots, seminal parameters appeared lower relative to historical values reported for zoo-managed ocelots, possibly related to reduced heterozygosity. In wild bobcats, seminal traits were inferior to those of ocelots but similar to reports for other zoo-managed Lynx species. In conclusion, detailed male reproductive traits have been characterized for the first time in wild, free-ranging ocelots and bobcats in southern Texas. Although UC allowed semen recovery for assessment of seminal traits in both species, EEJ produced higher quality samples in ocelots when applied after UC while also mitigating the adverse impact of urine contamination observed frequently with both collection methods.

Effects of Zinc (Zn) on the Growth of the Progeny from the Formation of Foetal Sperm through Parturition: A Review

The amount of zinc (Zn), another extremely important trace element, in human beings is substantial. Since the human body is unable to retain zinc, dietary intake of zinc is necessary for a number of bodily processes and metabolism. The testicles, epididymis, and prostate are the three main accessory sex glands, and normal development of these glands depends on the intake of Zn as it moves through the body. The early phases of the growth of germ cells and sperm development, the stages of sperm cell maturation and development, ejaculating the liquefaction the attachment of the male reproductive system and prostasomes, capacity, and fertilization are all important processes in which it participates. During ejaculation, the male reproductive system discharges higher zinc into the seminal fluid, which is important for the release and motility of sperm. The portion of Zn is essential during the gestational, labour, perinatal, and neonatal periods. During the maternity period, the average daily dietary intake of zinc in developing nations is 8–12 mg. Critical evidence has been used to discuss the lack of Zn and its effects. A thorough summary of the activities and roles played by Zn in effective fertilization has been provided. In the simplest terms, our current research highlights Zn's importance at every step of human reproduction, from spermatogenesis through birthing. Future research on reproductive biology now has a new avenue to explore thanks to the significance of zinc and its supplementation in in-vitro fertilization (IVF).

Cryopreservation Cooling Rate Impacts Post-Thaw Sperm Motility and Survival in Litoria booroolongensis.

The cryopreservation and storage of gametes (biobanking) can provide a long-term, low-cost option for the preservation of population genetic diversity and is particularly impactful when applied to manage selective breeding within conservation breeding programs (CBPs). This study aimed to develop a sperm cryopreservation protocol for the critically endangered Booroolong frog (Litoria booroolongensis) to capture founder genetics within the recently established (est. 2019) CBP for this species. Hormone-induced sperm release was achieved using established protocols, and spermic urine samples were collected over a 6-h period. Pooled spermic urine samples (n = 3 males) were divided equally between two cryoprotectant (CPA) treatments and diluted by 1:5 (sperm:CPA) with either 15% (v/v) dimethyl sulfoxide + 1% (w/v) sucrose in simplified amphibian Ringer's (SAR; CPAA) or 10% (v/v) dimethylformamide + 10% (w/v) trehalose dihydrate in SAR (CPAB). The samples were cryopreserved in 0.25 mL straws using either a programmable freezer (FrA) or an adapted dry shipper method (FrB). The thawed samples were activated via dilution in water and assessed for viability and motility using both manual assessment and computer-assisted sperm analysis (CASA; 0 h, 0.5 h post-thaw). Upon activation, the survival and recovery of motility (total motility, forward progression and velocity) of cryopreserved sperm suspensions were higher for sperm preserved using FrB than FrA, regardless of CPA composition. This work supports our long-term goal to pioneer the integration of biobanked cryopreserved sperm with population genetic management to maximize restoration program outcomes for Australian amphibian species.

Sperm allocation in relation to male–male aggression and courtship in an externally fertilizing fish, the medaka

Sperm production is physiologically costly; therefore, males that mate multiple times are expected to allocate sperm prudently to maximize their fitness. Sperm allocation theory has been widely tested in internally fertilizing animals; however, few studies have investigated whether males adjust the number of sperm released according to their premating experience and cues from rival males. Here, we investigated sperm allocation strategies in an externally fertilizing fish, the medaka, Oryzias latipes, in controlled conditions. We used three treatments: direct interaction with a rival (two males were allowed to interact directly with each other), visual cue only (presenting a male in a separate compartment) and no rivals (control). In the direct interaction treatment, two types of alternative male mating behaviour, simultaneous sperm release and postspawn sperm release, were observed. Premating experiences affected subsequent mating behaviour: males that were more aggressive and exhibited more prominent courtship behaviour mated females without any disturbance from the other males, whereas simultaneous sperm release frequently occurred when both males exhibited similar levels of aggression and courtship behaviours. We also counted the sperm released per mating from the water tank. There was no change in the number of sperm released in the presence of visual cues of competition or direct interaction with the other males before mating. However, males (focal and/or rival) released more sperm only when sperm competition was high, that is, when a rival male successfully disrupted mating before simultaneous sperm release. Our results indicate that medaka males may adjust the number of sperm released per mating in different mating environments.

Reproductive characteristics and gamete development of the soft coral Sclerophytum cf. heterospiculatum in Okinawa Island, Japan

AbstractSexual reproduction data are important to understand how organisms can replenish their populations and proliferate on coral reefs. Despite the importance of such data, the reproductive characteristics of most soft coral species are still unknown. Here, we examined the reproductive strategies of a species from the often‐dominant genus Sclerophytum in a coral reef on subtropical Okinawa Island, Japan. DNA barcoding and histological examinations of the tissues were conducted to confirm colony conspecificity and identify reproductive characteristics, respectively, between March 2020 and March 2021. Results indicated that the studied species, identified as Sclerophytum cf. heterospiculatum, exhibits gonochorism with longer oogenesis and shorter spermatogenesis. Female colonies produced immature oocytes throughout the year, with mature oocytes observed from late July to early September, and thus, extended spawning is likely characteristic of this species. In male colonies, spermatogenesis took place over ~5 months, with spermaries present from April through August. Mature spermaries were noted beginning in July and the inferred peak of sperm release was between late August and early September, which suggests that spermatogenesis duration was ~5 months. The largest mean oocyte and spermary sizes (628.45 ± 61.36 and 240.04 ± 49.49 μm, respectively) were both recorded in August. Gamete spawning presumably occurred during the summer season, which suggests seasonality in reproduction as influenced by changes in seawater temperature. However, the proximate cue for exact dates of spawning could be the lunar period because the inferred release of spawning materials seemed to occur between full moon and last‐quarter moon phases in both the months of August and September. The results of this study represent the first detailed report of reproductive characteristics of the genus Sclerophytum in Japan.

First babies conceived with Automated Intracytoplasmic Sperm Injection

Research questionCan an automated sperm injection robot perform Automated Intracytoplasmic Sperm Injection (ICSIA) for use in human IVF? DesignThe ICSIA robot automated the sperm injection procedure, including injection pipette advancement, zona pellucida and oolemma penetration with piezo pulses, and pipette removal after sperm release. The robot was first tested in mouse, hamster and rabbit oocytes, and subsequently using discarded human oocytes injected with microbeads. A small clinical pilot trial was conducted with donor oocytes to study the feasibility of the robot in a clinical setting. The ICSIA robot was controlled by engineers with no micromanipulation experience. Results were compared with those obtained with manual ICSI conducted by experienced embryologists. ResultsThe ICSIA robot demonstrated similar results to the manual procedure in the different animal models tested as well as in the pre-clinical validations conducted in discarded human oocytes. In the clinical validation, 13 out of 14 oocytes injected with ICSIA fertilized correctly versus 16 out of 18 in the manual control; eight developed into good-quality blastocysts versus 12 in the manual control; and four were diagnosed as chromosomally normal versus 10 euploid in the manual control. Three euploid blastocysts from the ICSIA robot group have been transferred into two recipients, which resulted in two singleton pregnancies and two babies born. ConclusionsThe ICSIA robot showed high proficiency in injecting animal and human oocytes when operated by inexperienced personnel. The preliminary results obtained in this first clinical pilot trial are within key performance indicators.

The carboxypeptidase B and carbonic anhydrase genes play a reproductive regulatory role during multiple matings in Ophraella communa.

Seminal fluid proteins (SFPs) are key factors in sexual reproduction and are transferred to females during mating with sperm. SFPs have a nutritional value because they protect and activate sperm storage and release to optimize fecundity. Multiple matings promote ovipositioning in several insect species. Therefore, insects may obtain more SFP through multiple matings to maximize reproduction, but this process has not yet been clearly confirmed. Here, the relationship between multiple matings and the SFPs in Ophraella communa (Coleoptera: Chrysomelidae), a biological control agent of the common ragweed Ambrosia artemisiifolia (Asterales: Asteraceae), was studied. Multiple matings significantly increased female fecundity and ovary egg deposition. Carboxypeptidase B (OcCpb) and carbonic anhydrase (OcCa) genes were identified as putative SFP genes in O. communa and they showed strong male-biased expression. Additionally, OcCpb and OcCa expression was upregulated in the bursa copulatrix of mating females compared to that in virgin females, but their expression gradually declined after copulation. Furthermore, OcCpb and OcCa knockdown in males led to a decrease in insect fecundity compared to that in the control. The reproductive tract of females mated with dsRNA-treated males was dissected and observed and, notably, the ovaries produced significantly fewer eggs. These data suggest that OcCpb and OcCa play regulatory roles during multiple matings in O. communa.

Es-β-CATENIN affects the hemolymph-testes barrier in Eriocheir sinensis by disrupting cell junctions and cytoskeleton

β-CATENIN is an evolutionarily conserved multifunctional molecule that maintains cell adhesion as a cell junction protein to safeguard the integrity of the mammalian blood-testes barrier, and also regulates cell proliferation and apoptosis as a key signaling molecule in the WNT/β-CATENIN signaling pathway. In the crustacean Eriocheir sinensis, Es-β-CATENIN has been shown to be involved in spermatogenesis, but the testes of E. sinensis have large and well-defined structural differences from those of mammals, and the impact of Es-β-CATENIN in them is still unknown. In the present study, we found that Es-β-CATENIN, Es-α-CATENIN and Es-ZO-1 interact differently in the testes of the crab compared to mammals. In addition, defective Es-β-CATENIN resulted in increased Es-α-CATENIN protein expression levels, distorted and deformed F-ACTIN, and disturbed localization of Es-α-CATENIN and Es-ZO-1, leading to loss of hemolymph-testes barrier integrity and impaired sperm release. In addition to this, we also performed the first molecular cloning and bioinformatics analysis of Es-AXIN in the WNT/β-CATENIN pathway to exclude the effect of the WNT/β-CATENIN pathway on the cytoskeleton. In conclusion, Es-β-CATENIN participates in maintaining the hemolymph-testes barrier in the spermatogenesis of E. sinensis.

LCRMP-1 is required for spermatogenesis and stabilises spermatid F-actin organization via the PI3K-Akt pathway

Long-form collapsin response mediator protein-1 (LCRMP-1) belongs to the CRMP family which comprises brain-enriched proteins responsible for axon guidance. However, its role in spermatogenesis remains unclear. Here we find that LCRMP-1 is abundantly expressed in the testis. To characterize its physiological function, we generate LCRMP-1-deficient mice (Lcrmp-1−/−). These mice exhibit aberrant spermiation with apoptotic spermatids, oligospermia, and accumulation of immature testicular cells, contributing to reduced fertility. In the seminiferous epithelial cycle, LCRMP-1 expression pattern varies in a stage-dependent manner. LCRMP-1 is highly expressed in spermatids during spermatogenesis and especially localized to the spermiation machinery during spermiation. Mechanistically, LCRMP-1 deficiency causes disorganized F-actin due to unbalanced signaling of F-actin dynamics through upregulated PI3K-Akt-mTOR signaling. In conclusion, LCRMP-1 maintains spermatogenesis homeostasis by modulating cytoskeleton remodeling for spermatozoa release.